C R Y S T A L L I Z A T I O N. volume 18 R E S E A R C H T O O L S

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hampton research • 34 journey • aliso viejo, ca 92656-3317 usa

telephone: 800-452-3899 or 949-425-1321 • fax: 949-425-1611

email: info@hrmail.com • website: www.hamptonresearch.com

v o l u m e

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C R Y S T A L L I Z A T I O N

S O L U T I O N S F O R C R Y S T A L G R O W T H

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c o n t a c t i n f o r m a t i o n

Business Hours: 7:00 a.m. to 5:00 p.m. Monday - Friday (Pacific Standard Time)

Phone: (800) 452-3899 or

(949) 425-1321

Fax: (949) 425-1611

Address: Hampton Research

34 Journey

Aliso Viejo, CA 92656 U.S.A.

c u s t o m e r s u p p o r t s e r v i c e s

Customer Service: Phone: (800) 452-3899 or

(949) 425-1321 Fax: (949) 425-1611 Email: info@hrmail.com Technical Support: Phone: (949) 425-1321

Fax: (949) 425-1611 Email: tech@hrmail.com

Website: www.hamptonresearch.com

On the cover: Crystals of Big-R protein of Xyllela fastidiosa.

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t a b l e o f c o n t e n t s

1 On the cover: Crystals of Big-R protein of Xyllela fastidiosa.

Produced in Brazilian Synchrotron Light Laboratory (LNLS),

2 - 33 s c r e e n s 34 - 37 c u s t o m s h o p c r y s t a l l i z a t i o n r e a g e n t s 38 - 59 o p t i m i z e™ c r y s t a l l i z a t i o n g r a d e r e a g e n t s 60 - 69 s t o c k o p t i o n s™ k i t s 70 - 105 c r y s t a l l i z a t i o n p l a t e s , h a r d w a r e & a c c e s s o r i e s 106 -115 t o o l s , s e e d i n g & r e s i n 116 - 135 c r y o c r y s t a l l o g r a p h y 136 - 145 c a p i l l a r y m o u n t s & s u p p l i e s 146 - 153 g o n i o m e t e r h e a d s & s u p p l i e s 154 - 157 x e n o n d e r i v a t i z a t i o n 158 - 163 l a b e l s a n d p e n s 164 - 173 b o o k s 174 - 179 p r o t e i n c r y s t a l l i z a t i o n s t a n d a r d s 180 - 267 c r y s t a l g r o w t h 1 0 1 268 - 275 i n d e x 276 - 277 g e n e r a l i n f o r m a t i o n P a G e s

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Reagent:

Crystal Screen Cryo Reagent 23 Mix equal amounts of

Glucose Isomerase and reagent. Vapor diffusion method. Mount crystal in

CryoLoop. Mosaicity may be a bit more in this

reagent and the unit cell will shrink a

s c r e e n s

Empire State Building-shaped crystal of neuron guidance receptor protein grown in Hampton Research Crystal Screen reagent 15.

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t a b l e o f c o n t e n t s

5 p c t™ p r e - c r y s t a l l i z a t i o n t e s t 6 - 7 i n d e x™ 8 - 9 c r y s t a l s c r e e n™ 10 - 11 p e g r x™ 12 - 13 p e g / i o n™ 14 - 15 g r i d s c r e e n s™ _•_{q u i k s c r e e n}™ 16 - 17 s a l t r x™ 18 - 19 m e m b f a c™_{• c r y s t a l s c r e e n l i t e}™ 20 - 21 n a t r i x™ 22 - 23 c r y s t a l s c r e e n c r y o™ 24 n u c l e i c a c i d m i n i s c r e e n™ 25 l o w i o n i c s t r e n g t h s c r e e n™ 26 - 27 s i l v e r b u l l e t s™ 28 a d d i t i v e s c r e e n™ 29 d e t e r g e n t s c r e e n™ 30 - 32 h e a v y a t o m k i t s™ P a G e s

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Crystals of kinase complex optimized from a Hampton Research Tacsimate based reagent.

Michelle Quiles, GlaxoSmithKline, RTP, North Carolina, USA.

Crystals of human mineralocorticoid receptor complexed with ligand. Grown using Hampton Salt Rx screen. Kevin P Madauss, GlaxoSmithkline, Research Triangle Park, North Carolina, USA.

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PCT Reagent A1/B1 Results PCT Reagent A2/B2 Results Action Heavy amorphous precipitate Heavy amorphous precipitate Dilute sample 1:1, repeat steps 1-7

Concentrate sample to half the original volume, repeat steps 1-7

Clear Clear

Perform screen Light granular precipitate Clear

Perform screen Clear Light granular precipitate

Perform screen Light granular precipitate Light granular precipitate

Heavy amorphous precipitate Clear Perform PCT with B1 & B2 / _{perform diagnostic testing} Clear Heavy amorphous precipitate Perform PCT with B1 & B2 / _{perform diagnostic testing}

Figure 1. Light Precipitate

Figure 2. Heavy Amorphous Precipitate n Determine the appropriate sample

concentra-tion for crystallizaconcentra-tion screening

n Conserve sample

n Optimize sample concentration prior to initial

screens

n Provide insight to sample homogeneity

a p p l i c a t i o n

f e a t u r e s

The PCT Pre-Crystallization Test is used to determine the appropriate sample concentration for crystallization screen-ing. Sample concentration is a signifi-cant crystallization variable. Samples too concentrated can result in amorphous precipitate, while samples too dilute can result in clear drops. Precipitate and

clear drops are typical crystallization screen results for reagent conditions which do not promote crystal-lization and are part of every crystalcrystal-lization screen. However, by optimizing protein concentration for the screen, the number of clear and precipitate results can often be reduced, which in turn results in more effi-cient sample utilization while at the same time enhancing the chances for crystallization. PCT can minimize or prevent situations where a screen results in an over abundance of precipitate or clear drops.

The PCT kit contains 4 unique, preformulated, sterile filtered reagents used to evaluate protein concentra-tion for crystallizaconcentra-tion screening. Initially, the sample protein is mixed with two of the reagents to determine if the protein concentration is appropriate for crystallization screening. If the protein is very sensitive to salt and polymer concentration, based on initial PCT results, the protein may be evaluated using a second set of PCT reagents. PCT results will then provide insight to either the appropriate sample concentration or indicate that other diagnostic testing such as native gel electrophoresis or dynamic light scattering should be performed to demonstrate sample homogeneity appropriate for crystallization.

Each PCT kit contains 4 unique reagents, 50 ml each. This kit is useful for researchers who already have crystallization plates and cover slides.

Each PCT kit (with plates) contains 4 unique reagents, 30 ml each, plus 24 well VDX™ Plates with sealant and plain cover slides. This kit is useful for labs without access to crystallization plates and cover slides.

References

1. Crystallization and x-ray diffraction analysis of human CLEC-2. Aleksandra A. Watson and Christopher A. O’Callaghan. Acta Cryst. (2005). F61, 1094–1096 Order Information

Each PCT kit contains 4 unique reagents. To order individual reagents, use Custom Shop™_catalog

number listed below. Refer to page 36 for further details.

Cat. No. Name Description Price

HR2-140 PCT 50 ml bottles (4 ea), plain cover slides (1 pk) $80.00

HR2-142 PCT (with Plates) 30 ml bottles (4 ea), plain cover slides (1 pk), $98.00

VDX Plates with sealant (5 ea)

HR2-940-** PCT Custom Shop 185 ml $138.00

** = reagent number A1-B2

d e s c r i p t i o n

screens

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Hampton Research • Phone: 800-452-3899 or 949-425-1321 • Fax: 949-425-1611 • www.hamptonresearch.com • Customer Service: info@hrmail.com • Technical Support: tech@hrmail.com

screens

n Primary, diverse reagent system

crystalliza-tion screen for proteins, complexes, peptides, nucleic acids, & water soluble small molecules

n Developed at Hampton Research

n A data-driven biased sparse matrix and grid

screen

n Screens classic, contemporary, & modern

crystallization reagents

n Samples pH 3 to 9

n Compatible with microbatch, vapor diffusion,

& liquid diffusion methods

n Specially formulated reagent zones: n Traditional salts versus pH n Neutralized organic acids n High [salt] with low [polymer] n High [polymer] with low [salt] n Low ionic strength versus pH n PEG & Salt versus pH n PEG & Salt

n Tube or Deep Well block format

f e a t u r e s

Index is designed as a 96 reagent crystallization screen that combines the strategies of the grid, sparse matrix, and incomplete factorial screening with traditional, con-temporary, and new crystallization reagent systems into a highly effective and efficient format.

Index, as the name implies, efficiently samples a series

of specially formulated reagent zones to identify which reagent class or classes and pH are effective in producing crystals or limiting sample solubility. Results from Index can be used to design optimization experiments and to identify follow on screens by reagent class. For example, positive results with salt based reagent in Index may be followed up with further screening using SaltRx or Grid Screen Salt. Success with polymer based reagents in Index may be followed up with further screening using PEGRx or PEG/Ion. Index utilizes a broad, yet refined portfolio of crystallization reagent systems. These include the following: (1) traditional salts such as Ammonium sulfate and Sodium chloride versus pH; (2) neutralized organic acids such as Sodium malonate and Tacsimate; (3) High salt concentration mixed with low polymer concentra-tion as well as high polymer concentraconcentra-tion mixed with low salt concentraconcentra-tion and; (4) Low ionic strength using polymers such as PEG, MPD, Pentaerythritols versus pH. These reagent systems are formulated across a sparse matrix and incomplete factorial of concentration ranges, sampling a pH range of 3 to 9. Index contains 96 unique reagents, 10 ml each.

Index HT contains 96 unique reagents in a single Deep Well block format.

Ready-to-use reagents are sterile filtered and formulated with ultra-pure Type 1 water, using the highest purity salts, polymers, organics and buffers.

Measured pH range of kit is 3 to 9 at 25°C Average measured pH of kit is 6.8 at 25°C Median measured pH of kit is 6.9 at 25°C

References

1. The advantages of using a modified microbatch method for rapid screening of protein crystallization conditions. A. D'Arcy, A. Mac Sweeney, M. Stihle and A. Haber. Acta Cryst. (2003). D59, 396-399.

2. Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of propionate kinase (TdcD) from Salmonella typhimurium. D. K. Simanshu and M. R. N. Murthy. Acta Cryst. (2005). F61, 52-55.

3. Preparation, crystallization and preliminary X-ray analysis of the methionine synthase (MetE) from Streptococcus mutans. T.-M. Fu, X.-Y. Zhang, L.-F. Li, Y.-H. Liang and X.-D. Su. Acta Cryst. (2006). F62, 984-985.

Crystals of a diabetes related protein.

Courtesy of Allan D’Arcy and Aengus Mac Sweeney.

Morphochem AG

s u c c e s s s t o r y

Order Information

Each Index kit contains 96 unique reagents. To order individual reagents, use Custom Shop™

cata-log number listed below. Refer to page 36 for further details.

HR2-144 Index 10 ml, tube format $570.00

HR2-134 Index HT 1 ml, Deep Well block format $185.00

HR2-944-** Index Custom Shop 185 ml $138.00

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screens

1. 0.1 M Citric Acid pH 3.5, 2.0 M Ammonium Sulfate 2. 0.1 M Sodium Acetate trihydrate pH 4.5, 2.0 M Ammonium Sulfate 3. 0.1 M Bis-Tris pH 5.5, 2.0 M Ammonium Sulfate

4. 0.1 M Bis-Tris pH 6.5, 2.0 M Ammonium Sulfate 5. 0.1 M HEPES pH 7.5, 2.0 M Ammonium Sulfate 6. 0.1 M Tris pH 8.5, 2.0 M Ammonium Sulfate 7. 0.1 M Citric Acid pH 3.5, 3.0 M Sodium Chloride 8. 0.1 M Sodium Acetate trihydrate pH 4.5, 3.0 M Sodium Chloride 9. 0.1 M Bis-Tris pH 5.5, 3.0 M Sodium Chloride

10. 0.1 M Bis-Tris pH 6.5, 3.0 M Sodium Chloride 11. 0.1 M HEPES pH 7.5, 3.0 M Sodium Chloride 12. 0.1 M Tris pH 8.5, 3.0 M Sodium Chloride 13. 0.1 M Bis-Tris pH 5.5, 0.3 M Magnesium Formate 14. 0.1 M Bis-Tris pH 6.5, 0.5 M Magnesium Formate 15. 0.1 M HEPES pH 7.5, 0.5 M Magnesium Formate 16. 0.1 M Tris pH 8.5, 0.3 M Magnesium Formate 17. 1.4 M Sodium/Potassium Phosphate pH 5.6 18. 1.4 M Sodium/Potassium Phosphate pH 6.9 19. 1.4 M Sodium/Potassium Phosphate pH 8.2 20. 0.1 M HEPES pH 7.5, 1.4 M tri-Sodium Citrate dihydrate 21. 1.8 M tri-Ammonium Citrate pH 7.0

22. 0.8 M Succinic Acid pH 7.0 23. 2.1 M DL-Malic Acid pH 7.0 24. 2.8 M Sodium Acetate trihydrate pH 7.0 25. 3.5 M Sodium Formate pH 7.0 26. 1.1 M di-Ammonium Tartrate pH 7.0 27. 2.4 M Sodium Malonate pH 7.0 28. 35% v/v Tacsimate pH 7.0 29. 60% v/v Tacsimate pH 7.0

30. 0.1 M Sodium Chloride, 0.1 M Bis-Tris pH 6.5, 1.5 M Ammonium Sulfate

31. 0.8 M Potassium Sodium Tartrate tetrahydrate, 0.1 M Tris pH 8.5, 0.5% w/v Polyethylene Glycol Monomethyl ether 5000 32. 1.0 M Ammonium Sulfate, 0.1 M Bis-Tris pH 5.5, 1% w/v Polyethylene Glycol 3350

33. 1.1 M Sodium Malonate pH 7.0, 0.1 M HEPES pH 7.0, 0.5% v/v Jeffamine ED-2001® Reagent pH 7.0 34. 1.0 M Succinic Acid pH 7.0, 0.1 M HEPES pH 7.0, 1% w/v Polyethylene Glycol Monomethyl ether 2000 35. 1.0 M Ammonium Sulfate, 0.1 M HEPES pH 7.0, 0.5% w/v Polyethylene Glycol 8000

36. 15% v/v Tacsimate pH 7.0, 0.1 M HEPES pH 7.0, 2% w/v Polyethylene Glycol 3350 37. 25% w/v Polyethylene Glycol 1500

38. 0.1 M HEPES pH 7.0, 30% v/v Jeffamine M-600® Reagent pH 7.0 39. 0.1 M HEPES pH 7.0, 30% v/v Jeffamine ED-2001® Reagent pH 7.0 40. 0.1 M Citric Acid pH 3.5, 25% w/v Polyethylene Glycol 3350 41. 0.1 M Sodium Acetate trihydrate pH 4.5, 25% w/v Polyethylene Glycol 3350 42. 0.1 M Bis-Tris pH 5.5, 25% w/v Polyethylene Glycol 3350

43. 0.1 M Bis-Tris pH 6.5, 25% w/v Polyethylene Glycol 3350 44. 0.1 M HEPES pH 7.5, 25% w/v Polyethylene Glycol 3350 45. 0.1 M Tris pH 8.5, 25% w/v Polyethylene Glycol 3350

46. 0.1 M Bis-Tris pH 6.5, 20% w/v Polyethylene Glycol Monomethyl ether 5000 47. 0.1 M Bis-Tris pH 6.5, 28% w/v Polyethylene Glycol Monomethyl ether 2000 48. 0.2 M Calcium Chloride dihydrate, 0.1 M Bis-Tris pH 5.5, 45% v/v 2-Methyl-2,4-pentanediol 49. 0.2 M Calcium Chloride dihydrate, 0.1 M Bis-Tris pH 6.5, 45% v/v 2-Methyl-2,4-pentanediol 50. 0.2 M Ammonium Acetate, 0.1 M Bis-Tris pH 5.5, 45% v/v 2-Methyl-2,4-pentanediol 51. 0.2 M Ammonium Acetate, 0.1 M Bis-Tris pH 6.5, 45% v/v 2-Methyl-2,4-pentanediol 52. 0.2 M Ammonium Acetate, 0.1 M HEPES pH 7.5, 45% v/v 2-Methyl-2,4-pentanediol 53. 0.2 M Ammonium Acetate, 0.1 M Tris pH 8.5, 45% v/v 2-Methyl-2,4-pentanediol

54. 0.05 M Calcium Chloride dihydrate, 0.1 M Bis-Tris pH 6.5, 30% v/v Polyethylene Glycol Monomethyl ether 550 55. 0.05 M Magnesium Chloride hexahydrate, 0.1 M HEPES pH 7.5, 30% v/v Polyethylene Glycol Monomethyl ether 550 56. 0.2 M Potassium Chloride, 0.05 M HEPES pH 7.5, 35% v/v Pentaerythritol Propoxylate (5/4 PO/OH)

57. 0.05 M Ammonium Sulfate, 0.05 M Bis-Tris pH 6.5, 30% v/v Pentaerythritol Ethoxylate (15/4 EO/OH) 58. 0.1 M Bis-Tris pH 6.5, 45% v/v Polypropylene Glycol P 400

59. 0.02 M Magnesium Chloride hexahydrate, 0.1 M HEPES pH 7.5, 22% w/v Polyacrylic Acid 5100 Sodium salt 60. 0.01 M Cobalt Chloride hexahydrate, 0.1 M Tris pH 8.5, 20% w/v Polyvinylpyrrolidone K15

61. 0.2 M Proline, 0.1 M HEPES pH 7.5, 10% w/v Polyethylene Glycol 3350

62. 0.2 M Trimethylamine N-oxide dihydrate, 0.1 M Tris pH 8.5, 20% w/v Polyethylene Glycol Monomethyl ether 2000 63. 5% v/v Tacsimate pH 7.0, 0.1 M HEPES pH 7.0, 10% w/v Polyethylene Glycol Monomethyl ether 5000

64. 0.005 M Cobalt Chloride hexahydrate, 0.005 M Nickel (II) Chloride hexahydrate, 0.005 M Cadmium Chloride dihydrate, 0.005 M Magnesium Chloride hexahydrate, 0.1 M HEPES pH 7.5, 12% w/v Polyethylene Glycol 3350

65. 0.1 M Ammonium Acetate, 0.1 M Bis-Tris pH 5.5, 17% w/v Polyethylene Glycol 10,000 66. 0.2 M Ammonium Sulfate, 0.1 M Bis-Tris pH 5.5, 25% w/v Polyethylene Glycol 3350 67. 0.2 M Ammonium Sulfate, 0.1 M Bis-Tris pH 6.5, 25% w/v Polyethylene Glycol 3350 68. 0.2 M Ammonium Sulfate, 0.1 M HEPES pH 7.5, 25% w/v Polyethylene Glycol 3350 69. 0.2 M Ammonium Sulfate, 0.1 M Tris pH 8.5, 25% w/v Polyethylene Glycol 3350 70. 0.2 M Sodium Chloride, 0.1 M Bis-Tris pH 5.5, 25% w/v Polyethylene Glycol 3350 71. 0.2 M Sodium Chloride, 0.1 M Bis-Tris pH 6.5, 25% w/v Polyethylene Glycol 3350 72. 0.2 M Sodium Chloride, 0.1 M HEPES pH 7.5, 25% w/v Polyethylene Glycol 3350 73. 0.2 M Sodium Chloride, 0.1 M Tris pH 8.5, 25% w/v Polyethylene Glycol 3350 74. 0.2 M Lithium Sulfate monohydrate, 0.1 M Bis-Tris pH 5.5, 25% w/v Polyethylene Glycol 3350 75. 0.2 M Lithium Sulfate monohydrate, 0.1 M Bis-Tris pH 6.5, 25% w/v Polyethylene Glycol 3350 76. 0.2 M Lithium Sulfate monohydrate, 0.1 M HEPES pH 7.5, 25% w/v Polyethylene Glycol 3350 77. 0.2 M Lithium Sulfate monohydrate, 0.1 M Tris pH 8.5, 25% w/v Polyethylene Glycol 3350 78. 0.2 M Ammonium Acetate, 0.1 M Bis-Tris pH 5.5, 25% w/v Polyethylene Glycol 3350 79. 0.2 M Ammonium Acetate, 0.1 M Bis-Tris pH 6.5, 25% w/v Polyethylene Glycol 3350 80. 0.2 M Ammonium Acetate, 0.1 M HEPES pH 7.5, 25% w/v Polyethylene Glycol 3350 81. 0.2 M Ammonium Acetate, 0.1 M Tris pH 8.5, 25% w/v Polyethylene Glycol 3350

82. 0.2 M Magnesium Chloride hexahydrate, 0.1 M Bis-Tris pH 5.5, 25% w/v Polyethylene Glycol 3350 83. 0.2 M Magnesium Chloride hexahydrate, 0.1 M Bis-Tris pH 6.5, 25% w/v Polyethylene Glycol 3350 84. 0.2 M Magnesium Chloride hexahydrate, 0.1 M HEPES pH 7.5, 25% w/v Polyethylene Glycol 3350 85. 0.2 M Magnesium Chloride hexahydrate, 0.1 M Tris HCl pH 8.5, 25% w/v Polyethylene Glycol 3350 86. 0.2 M Potassium Sodium Tartrate tetrahydrate, 20% w/v Polyethylene Glycol 3350

87. 0.2 M Sodium Malonate pH 7.0, 20% w/v Polyethylene Glycol 3350 88. 0.2 M tri-Ammonium Citrate pH 7.0, 20% w/v Polyethylene Glycol 3350 89. 0.1 M Succinic Acid pH 7.0, 15% w/v Polyethylene Glycol 3350 90. 0.2 M Sodium Formate, 20% w/v Polyethylene Glycol 3350 91. 0.15 M DL-Malic Acid pH 7.0, 20% w/v Polyethylene Glycol 3350 92. 0.1 M Magnesium Formate, 15% w/v Polyethylene Glycol 3350 93. 0.05 M Zinc Acetate dihydrate, 20% w/v Polyethylene Glycol 3350 94. 0.2 M tri-Sodium Citrate dihydrate, 20% w/v Polyethylene Glycol 3350 95. 0.1 M Potassium Thiocyanate, 30% w/v Polyethylene Glycol Monomethyl ether 2000 96. 0.15 M Potassium Bromide, 30% w/v Polyethylene Glycol Monomethyl ether 2000

i n d e x™ f o r m u l a t i o n A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12 B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12 B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 Range from 3 to 9 pH h i g h [ p o l y m e r ] / l o w [ s a l t ] n e u t r a l i z e d o r g a n i c a c i d s t r a d i t i o n a l s a l t s p o l y m e r / s a l t

i n d e x

f a c t o r s

h i g h [ s a l t ] / l o w [ p o l y m e r ] p o l y m e r / s a l t / p H l o w i o n i c s t r e n g t h

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n Primary screen for proteins, soluble peptides,

nucleic acids, & water soluble small molecules

n Sparse matrix additive screen

n The original sparse matrix screen

n Sparse matrix formula efficiently samples

salts, polymers, organics, & pH

n Proven effective with more than 1,000

biological macromolecules

n Tube or DeepWell block format

f e a t u r e s

The Crystal Screen and Crystal Screen 2 reagent kits are designed to provide a highly effective and rapid screening method for the crystallization of macromolecules. The screens are simple and practical for finding initial crystallization condi-tions. The initial crystallization conditions for more than 1,000 proteins, peptides, oligonucle-otides, and small molecules have been deter-mined using Crystal Screen.

A highly effective approach to overcome the exhaustive search for suitable crystallization conditions is the use of a sparse matrix method of trial conditions that is biased and selected from known crystallization conditions for macromolecules. The formulation utilized in Crystal Screen and Crystal Screen 2 evaluates 96 unique mixtures of pH, salts, polymers and organics, and their ability to promote crystal growth. Crystal Screen contains 50 unique reagents, 10 ml each and is based on the sparse matrix formulation first described by Jancarik and Kim in 1991.

Crystal Screen 2, an extension of Crystal Screen, contains 48 unique reagents, 10 ml each and is based on the formulation first described by Cudney et al in 1994.

Crystal Screen HT contains 1 ml each of reagents 1-48 from Crystal Screen and all 48 reagents from Crystal Screen 2 in a single Deep Well block format.

References

1. Jancarik, J. & Kim, S.H. J. Appl. Cryst. 24, 409-411, (1991).

2. Expression, purification, crystallization and preliminary X-ray analysis of two arginine-biosynthetic enzymes from Mycobacterium tuberculosis. F. Moradian, C. Garen, L. Cherney, M. Cherney and M. N. G. James. Acta Cryst. (2006). F62, 986-988.

d e s c r i p t i o n

Crystal of gp120, the exterior envelope glycoprotein of type 1 HIV. Preliminary crystallization conditions obtained using Crystal Screen from Hampton Research.

Courtesy of P.D. Kwong1_{, R. Wyatt}2_{, E. Desjardins}2_, J. Robinson3_{, F.C. Culp}4_{, B.D. Hellmig}4_{, R.W. Sweet}4_, J. Sodroski2_{, and W.A. Hendrickson}1,5_.

1_{Columbia University,}2_{Dana-Farber Cancer Institute,}3_Tulane University School of Medicine, 4_{GlaxoSmithKline ,}5 HHMI-Columbia University

s u c c e s s s t o r y

screens

Each Crystal Screen kit contains 50 unique reagents. Each Crystal Screen 2 kit contains 48 unique reagents. To order individual reagents, use Custom Shop™_{catalog numbers listed below. Refer to}

page 36 for further details.

HR2-110 Crystal Screen 10 ml, tube format $285.00

HR2-112 Crystal Screen 2 10 ml, tube format $285.00

HR2-130 Crystal Screen HT 1 ml, Deep Well block format $185.00

HR2-910-** Crystal Screen Custom Shop 185 ml $138.00

HR2-912-** Crystal Screen 2 Custom Shop 185 ml $138.00

** = reagent number 1-50 (for Crystal Screen) ** = reagent number 1-48 (for Crystal Screen 2)

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c r y s ta l s c r e e n

™

_{• C r y s Ta l s C r e e n 2}

™

_{• C r y s Ta l s C r e e n}

_{H T}

™ screens A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12 B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 c r y s t a l s c r e e n ™ f o r m u l a t i o n

1. 0.02 M Calcium chloride dihydrate, 0.1 M Sodium acetate trihydrate pH 4.6, 30% v/v (+/-)-2-Methyl-2,4-pentanediol 2. 0.4 M Potassium sodium tartrate tetrahydrate

3. 0.4 M Ammonium phosphate monobasic

4. 0.1 M TRIS hydrochloride pH 8.5, 2.0 M Ammonium sulfate

5. 0.2 M Sodium citrate tribasic dihydrate, 0.1 M HEPES sodium pH 7.5, 30% v/v (+/-)-2-Methyl-2,4-pentanediol 6. 0.2 M Magnesium chloride hexahydrate, 0.1 M TRIS hydrochloride pH 8.5, 30% w/v Polyethylene glycol 4,000 7. 0.1 M Sodium cacodylate trihydrate pH 6.5, 1.4 M Sodium acetate trihydrate

8. 0.2 M Sodium citrate tribasic dihydrate, 0.1 M Sodium cacodylate trihydrate pH 6.5, 30% v/v 2-Propanol 9. 0.2 M Ammonium acetate, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 30% w/v Polyethylene glycol 4,000 10. 0.2 M Ammonium acetate, 0.1 M Sodium acetate trihydrate pH 4.6, 30% w/v Polyethylene glycol 4,000 11. 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 1.0 M Ammonium phosphate monobasic 12. 0.2 M Magnesium chloride hexahydrate, 0.1 M HEPES sodium pH 7.5, 30% v/v 2-Propanol 13. 0.2 M Sodium citrate tribasic dihydrate, 0.1 M TRIS hydrochloride pH 8.5, 30% v/v Polyethylene glycol 400 14. 0.2 M Calcium chloride dihydrate, 0.1 M HEPES sodium pH 7.5, 28% v/v Polyethylene glycol 400 15. 0.2 M Ammonium sulfate, 0.1 M Sodium cacodylate trihydrate pH 6.5, 30% w/v Polyethylene glycol 8,000 16. 0.1 M HEPES sodium pH 7.5, 1.5 M Lithium sulfate monohydrate

17. 0.2 M Lithium sulfate monohydrate, 0.1 M TRIS hydrochloride pH 8.5, 30% w/v Polyethylene glycol 4,000 18. 0.2 M Magnesium acetate tetrahydrate, 0.1 M Sodium cacodylate trihydrate pH 6.5, 20% w/v Polyethylene glycol 8,000 19. 0.2 M Ammonium acetate, 0.1 M TRIS hydrochloride pH 8.5, 30% v/v 2-Propanol

20. 0.2 M Ammonium sulfate, 0.1 M Sodium acetate trihydrate pH 4.6, 25% w/v Polyethylene glycol 4,000

21. 0.2 M Magnesium acetate tetrahydrate, 0.1 M Sodium cacodylate trihydrate pH 6.5, 30% v/v (+/-)-2-Methyl-2,4-pentanediol 22. 0.2 M Sodium acetate trihydrate, 0.1 M TRIS hydrochloride pH 8.5, 30% w/v Polyethylene glycol 4,000

23. 0.2 M Magnesium chloride hexahydrate, 0.1 M HEPES sodium pH 7.5, 30% v/v Polyethylene glycol 400 24. 0.2 M Calcium chloride dihydrate, 0.1 M Sodium acetate trihydrate pH 4.6, 20% v/v 2-Propanol 25. 0.1 M Imidazole pH 6.5, 1.0 M Sodium acetate trihydrate

26. 0.2 M Ammonium acetate, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 30% v/v (+/-)-2-Methyl-2,4-pentanediol 27. 0.2 M Sodium citrate tribasic dihydrate, 0.1 M HEPES sodium pH 7.5, 20% v/v 2-Propanol

28. 0.2 M Sodium acetate trihydrate, 0.1 M Sodium cacodylate trihydrate pH 6.5, 30% w/v Polyethylene glycol 8,000 29. 0.1 M HEPES sodium pH 7.5, 0.8 M Potassium sodium tartrate tetrahydrate

30. 0.2 M Ammonium sulfate, 30% w/v Polyethylene glycol 8,000 31. 0.2 M Ammonium sulfate, 30% w/v Polyethylene glycol 4,000 32. 2.0 M Ammonium sulfate

33. 4.0 M Sodium formate

34. 0.1 M Sodium acetate trihydrate pH 4.6, 2.0 M Sodium formate

35. 0.1 M HEPES sodium pH 7.5, 0.8 M Sodium phosphate monobasic monohydrate, 0.8 M Potassium phosphate monobasic 36. 0.1 M TRIS hydrochloride pH 8.5, 8% w/v Polyethylene glycol 8,000

37. 0.1 M Sodium acetate trihydrate pH 4.6, 8% w/v Polyethylene glycol 4,000 38. 0.1 M HEPES sodium pH 7.5, 1.4 M Sodium citrate tribasic dihydrate

39. 0.1 M HEPES sodium pH 7.5, 2% v/v Polyethylene glycol 400, 2.0 M Ammonium sulfate 40. 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 20% v/v 2-Propanol, 20% w/v Polyethylene glycol 4,000 41. 0.1 M HEPES sodium pH 7.5, 10% v/v 2-Propanol, 20% w/v Polyethylene glycol 4,000

42. 0.05 M Potassium phosphate monobasic, 20% w/v Polyethylene glycol 8,000 43. 30% w/v Polyethylene glycol 1,500

44. 0.2 M Magnesium formate dihydrate

45. 0.2 M Zinc acetate dihydrate, 0.1 M Sodium cacodylate trihydrate pH 6.5, 18% w/v Polyethylene glycol 8,000 46. 0.2 M Calcium acetate hydrate, 0.1 M Sodium cacodylate trihydrate pH 6.5 , 18% w/v Polyethylene glycol 8,000 47. 0.1 M Sodium acetate trihydrate pH 4.6, 2.0 M Ammonium sulfate

48. 0.1 M TRIS hydrochloride pH 8.5, 2.0 M Ammonium phosphate monobasic 49. 1.0 M Lithium sulfate monohydrate, 2% w/v Polyethylene glycol 8,000 50. 0.5 M Lithium sulfate monohydrate, 15% w/v Polyethylene glycol 8,000

1. 2.0 M Sodium chloride, 10% w/v Polyethylene glycol 6,000

2. 0.5 M Sodium chloride, 0.01 M Magnesium chloride hexahydrate, 0.01 M Hexadecyltrimethylammonium bromide 3. 25% v/v Ethylene glycol

4. 35% v/v 1,4-Dioxane

5. 2.0 M Ammonium sulfate, 5% v/v 2-Propanol 6. 1.0 M Imidazole pH 7.0

7. 10% w/v Polyethylene glycol 1,000, 10% w/v Polyethylene glycol 8,000 8. 1.5 M Sodium chloride, 10% v/v Ethanol

9. 0.1 M Sodium acetate trihydrate pH 4.6, 2.0 M Sodium chloride

10. 0.2 M Sodium chloride, 0.1 M Sodium acetate trihydrate pH 4.6, 30% v/v (+/-)-2-Methyl-2,4-pentanediol 11. 0.01 M Cobalt(II) chloride hexahydrate, 0.1 M Sodium acetate trihydrate pH 4.6, 1.0 M 1,6-Hexanediol 12. 0.1 M Cadmium chloride hydrate, 0.1 M Sodium acetate trihydrate pH 4.6, 30% v/v Polyethylene glycol 400 13. 0.2 M Ammonium sulfate, 0.1 M Sodium acetate trihydrate pH 4.6, 30% w/v Polyethylene glycol monomethyl ether 2,000 14. 0.2 M Potassium sodium tartrate tetrahydrate, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 2.0 M Ammonium sulfate 15. 0.5 M Ammonium sulfate, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 1.0 M Lithium sulfate monohydrate 16. 0.5 M Sodium chloride, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 2% v/v Ethylene imine polymer 17. 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 35% v/v tert-Butanol

18. 0.01 M Iron(III) chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 10% v/v Jeffamine ® M-600 ®

19. 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 2.5 M 1,6-Hexanediol 20. 0.1 M MES monohydrate pH 6.5, 1.6 M Magnesium sulfate heptahydrate

21. 0.1 M Sodium phosphate monobasic monohydrate, 0.1 M Potassium phosphate monobasic, 0.1 M MES monohydrate pH 6.5, 2.0 M Sodium chloride

22. 0.1 M MES monohydrate pH 6.5, 12% w/v Polyethylene glycol 20,000 23. 1.6 M Ammonium sulfate, 0.1 M MES monohydrate pH 6.5, 10% v/v 1,4-Dioxane 24. 0.05 M Cesium chloride, 0.1 M MES monohydrate pH 6.5, 30% v/v Jeffamine ® M-600 ®

25. 0.01 M Cobalt(II) chloride hexahydrate, 0.1 M MES monohydrate pH 6.5, 1.8 M Ammonium sulfate 26. 0.2 M Ammonium sulfate, 0.1 M MES monohydrate pH 6.5, 30% w/v Polyethylene glycol monomethyl ether 5,000 27. 0.01 M Zinc sulfate heptahydrate, 0.1 M MES monohydrate pH 6.5, 25% v/v Polyethylene glycol monomethyl ether 550 28. 1.6 M Sodium citrate tribasic dihydrate pH 6.5

29. 0.5 M Ammonium sulfate, 0.1 M HEPES pH 7.5, 30% v/v (+/-)-2-Methyl-2,4-pentanediol 30. 0.1 M HEPES pH 7.5, 10% w/v Polyethylene glycol 6,000, 5% v/v (+/-)-2-Methyl-2,4-pentanediol 31. 0.1 M HEPES pH 7.5, 20% v/v Jeffamine ® M-600 ®

32. 0.1 M Sodium chloride, 0.1 M HEPES pH 7.5, 1.6 M Ammonium sulfate 33. 0.1 M HEPES pH 7.5, 2.0 M Ammonium formate

34. 0.05 M Cadmium sulfate hydrate, 0.1 M HEPES pH 7.5, 1.0 M Sodium acetate trihydrate 35. 0.1 M HEPES pH 7.5, 70% v/v (+/-)-2-Methyl-2,4-pentanediol

36. 0.1 M HEPES pH 7.5, 4.3 M Sodium chloride

37. 0.1 M HEPES pH 7.5, 10% w/v Polyethylene glycol 8,000, 8% v/v Ethylene glycol 38. 0.1 M HEPES pH 7.5, 20% w/v Polyethylene glycol 10,000

39. 0.2 M Magnesium chloride hexahydrate, 0.1 M Tris pH 8.5, 3.4 M 1,6-Hexanediol 40. 0.1 M Tris pH 8.5, 25% v/v tert-Butanol

41. 0.01 M Nickel(II) chloride hexahydrate, 0.1 M Tris pH 8.5, 1.0 M Lithium sulfate monohydrate 42. 1.5 M Ammonium sulfate, 0.1 M Tris pH 8.5, 12% v/v Glycerol

43. 0.2 M Ammonium phosphate monobasic, 0.1 M Tris pH 8.5, 50% v/v (+/-)-2-Methyl-2,4-pentanediol 44. 0.1 M Tris pH 8.5, 20% v/v Ethanol

45. 0.01 M Nickel(II) chloride hexahydrate, 0.1 M Tris pH 8.5, 20% w/v Polyethylene glycol monomethyl ether 2,000 46. 0.1 M Sodium chloride, 0.1 M BICINE pH 9.0, 20% v/v Polyethylene glycol monomethyl ether 550 47. 0.1 M BICINE pH 9.0, 2.0 M Magnesium chloride hexahydrate

48. 0.1 M BICINE pH 9.0, 2% v/v 1,4-Dioxane, 10% w/v Polyethylene glycol 20,000

c r y s t a l s c r e e n 2 ™ f o r m u l a t i o n Range from 4.0 to 9.0 1,4-Dioxane Ethanol 2-Propanol tert-Butanol 1,6-Hexanediol (+/-)-2-Methyl-2,4-pentanediol Ethylene glycol Glycerol

Ethylene imine polymer Jeffamine ® M-600 ® Polyethylene glycol 400 Polyethylene glycol 1,000 Polyethylene glycol 6,000 Polyethylene glycol 8,000 Polyethylene glycol 10,000 Polyethylene glycol 20,000 Polyethylene glycol MME 550 Polyethylene glycol MME 2,000 Polyethylene glycol MME 5,000 BICINE HEPES Imidazole MES Sodium acetate Sodium citrate TRIS hydrochloride o r g a n i c pH n o n - v o l a t i l e o r g a n i c s a l t p o l y m e r b u f f e r Ammonium phosphate Potassium phosphate Sodium phosphate Ammonium sulfate Cadmium sulfate Lithium sulfate Magnesium sulfate Zinc sulfate K/Na tartrate Hexadecyltrimethylammonium bromide Sodium acetate Cadmium chloride Cesium chloride Cobalt(II) chloride Iron(III) chloride Magnesium chloride Nickel(II) chloride Sodium chloride Sodium citrate Ammonium formate p o l y m e r Range from 4.0 to 9.0 2-Propanol (+/-)-2-Methyl-2,4-pentanediol Polyethylene glycol 400 Polyethylene glycol 1,500 Polyethylene glycol 4,000 Polyethylene glycol 8,000 Ammonium acetate Calcium acetate Magnesium acetate Sodium acetate Zinc acetate Calcium chloride Magnesium chloride Sodium citrate Magnesium formate Sodium formate Ammonium phosphate Potassium phosphate Sodium phosphate Ammonium sulfate Lithium sulfate K/Na tartrate o r g a n i c pH n o n - v o l a t i l e o r g a n i c s a l t HEPES sodium Imidazole Sodium acetate Sodium cacodylate Sodium citrate TRIS hydrochloride b u f f e r

crystal screen

factors

crystal screen

2 factors

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n Primary and secondary, polymer and pH

based crystallization screen for biological macromolecules

n PEGRx 1 primary screen variables are

polymer type (16 different polymers), polymer molecular weight, pH and low ionic strength.

n PEGRx 2 primary screen variables are

polymer type (13 different polymers), polymer molecular weight, pH and secondary reagents which include additives, salts, volatile organics and polyols.

n PEGRx HT combines PEGRx 1 and PEGRx 2

in a single 96 Deep Well block

n pH range 3.5 - 9, using 10 different buffer

systems

n Polymer molecular weight range 200 to

20,000

f e a t u r e s

PEGRx is a primary and secondary, polymer and pH based crystallization screen devel-oped at Hampton Research. It is

designed to evaluate polymer based crystal-lization reagents and pH in low (PEGRx 1) to medium ionic strength (PEGRx 2). It is also designed for use as a secondary or optimiza-tion screen to follow the Hampton Research Index screen when polymer based reagents produce hits and interesting solubility leads.

PEGRx 1 is a crystallization reagent kit designed to evaluate an array of polymers of varying molecu-lar weight in a low ionic strength environment versus a wide range of pH. Polymer reagents include Polyethylene glycols, Polyethylene glycol monomethylethers, and Jeffamines. The molecular weight range between 200 and 20,000 is evaluated in a low ionic strength formulation. Ten different buffers are used to span the range of pH between 3.5 and 9. The primary screen variables are polymer type, polymer molecular weight, pH and low ionic strength.

PEGRx 2 is a crystallization reagent kit designed to evaluate an array of polymers of varying molecular weight in a medium ionic strength environment in the presence of additives, salts, volatile organics and polyols versus a wide range of pH. Polymer reagents include Polyethylene glycols and Polyethylene gly-col monomethylethers. The polymer molecular weight range between 200 and 20,000 is evaluated in a medium ionic strength formulation. Ten different buffers are used to span the range of pH between 3.5 and 9. The primary screen variables are polymer type, polymer molecular weight, pH and secondary reagents which include additives, salts, volatile organics and polyols.

PEGRx 1 contains 48 unique reagents, 10 ml each. PEGRx 2 contains 48 unique reagents, 10 ml each.

PEGRx HT contains 1 ml of each reagent from PEGRx 1 and PEGRx 2 in a single Deep Well block format. Ready-to-use reagents are sterile filtered and formulated with ultra-pure Type 1 water, using the highest purity salts, polymers, organics and buffers.

PEGRx 1 and PEGRx 2 kits each contain 48 unique reagents. To order individual reagents, use Custom Shop™_{catalog numbers listed below. Refer to page 36 for further details.}

HR2-082 PEGRx 1 10 ml, tube format $285.00

HR2-084 PEGRx 2 10 ml, tube format $285.00

HR2-086 PEGRx HT 1 ml, Deep Well block format $185.00

HR2-982-** PEGRx 1 Custom Shop 185 ml $138.00

HR2-984-** PEGRx 2 Custom Shop 185 ml $138.00

** = reagent number 1-48

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screens A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12 B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 P e G r x 1 ™ f o r m u l a t i o n

1. 0.1 M Citric acid pH 3.5, 34% v/v Polyethylene glycol 200

2. 0.1 M Sodium citrate tribasic dihydrate pH 5.5, 38% v/v Polyethylene glycol 200 3 0.1 M HEPES pH 7.5, 42% v/v Polyethylene glycol 200

4. 0.1 M Sodium acetate trihydrate pH 4.5, 30% v/v Polyethylene glycol 300 5. 0.1 M BIS-TRIS pH 6.5, 25% v/v Polyethylene glycol 300 6. 0.1 M BICINE pH 8.5, 20% v/v Polyethylene glycol 300

7. 0.1 M Sodium acetate trihydrate pH 4.0, 15% v/v Polyethylene glycol 400 8. 0.1 M MES monohydrate pH 6.0, 22% v/v Polyethylene glycol 400 9. 0.1 M Tris pH 8.0, 30% v/v Polyethylene glycol 400

10. 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 30% v/v Polyethylene glycol monomethyl ether 550 11. 0.1 M Imidazole pH 7.0, 25% v/v Polyethylene glycol monomethyl ether 550

12. 0.1 M BIS-TRIS propane pH 9.0, 20% v/v Polyethylene glycol monomethyl ether 550 13. 0.1 M Sodium acetate trihydrate pH 4.0, 10% v/v Jeffamine ® M-600 ® pH 7.0 14. 0.1 M MES monohydrate pH 6.0, 20% v/v Jeffamine ® M-600 ® pH 7.0 15. 0.1 M Tris pH 8.0, 30% v/v Jeffamine ® M-600 ® pH 7.0 16. 0.1 M Citric acid pH 3.5, 14% w/v Polyethylene glycol 1,000

17. 0.1 M Sodium citrate tribasic dihydrate pH 5.5, 22% w/v Polyethylene glycol 1,000 18. 0.1 M HEPES pH 7.5, 30% w/v Polyethylene glycol 1,000

19. 0.1 M Sodium acetate trihydrate pH 4.5, 30% w/v Polyethylene glycol 1,500 20. 0.1 M BIS-TRIS pH 6.5, 20% w/v Polyethylene glycol 1,500 21. 0.1 M BICINE pH 8.5, 15% w/v Polyethylene glycol 1,500

22. 0.1 M Sodium acetate trihydrate pH 4.0, 10% w/v Polyethylene glycol monomethyl ether 2,000 23. 0.1 M MES monohydrate pH 6.0, 20% w/v Polyethylene glycol monomethyl ether 2,000 24. 0.1 M Tris pH 8.0, 30% w/v Polyethylene glycol monomethyl ether 2,000 25. 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 30% v/v Jeffamine ® ED-2001 pH 7.0 26. 0.1 M Imidazole pH 7.0, 20% v/v Jeffamine ® ED-2001 pH 7.0

27. 0.1 M BIS-TRIS propane pH 9.0, 10% v/v Jeffamine ® ED-2001 pH 7.0 28. 0.1 M Citric acid pH 3.5, 25% w/v Polyethylene glycol 3,350

31. 0.1 M Sodium acetate trihydrate pH 4.0, 10% w/v Polyethylene glycol 4,000 32. 0.1 M MES monohydrate pH 6.0, 14% w/v Polyethylene glycol 4,000 33. 0.1 M Tris pH 8.0, 28% w/v Polyethylene glycol 4,000

34. 0.1 M Sodium acetate trihydrate pH 4.5, 30% w/v Polyethylene glycol monomethyl ether 5,000 35. 0.1 M BIS-TRIS pH 6.5, 20% w/v Polyethylene glycol monomethyl ether 5,000 36. 0.1 M BICINE pH 8.5, 8% w/v Polyethylene glycol monomethyl ether 5,000 37. 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 10% w/v Polyethylene glycol 6,000 38. 0.1 M Imidazole pH 7.0, 20% w/v Polyethylene glycol 6,000

39. 0.1 M BIS-TRIS propane pH 9.0, 30% w/v Polyethylene glycol 6,000 40. 0.1 M Citric acid pH 3.5, 28% w/v Polyethylene glycol 8,000

43. 0.1 M Sodium acetate trihydrate pH 4.5, 10% w/v Polyethylene glycol 10,000 44. 0.1 M BIS-TRIS pH 6.5, 16% w/v Polyethylene glycol 10,000 45. 0.1 M BICINE pH 8.5, 20% w/v Polyethylene glycol 10,000

46. 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 18% w/v Polyethylene glycol 20,000 47. 0.1 M Imidazole pH 7.0, 12% w/v Polyethylene glycol 20,000

48. 0.1 M BIS-TRIS propane pH 9.0, 8% w/v Polyethylene glycol 20,000

1. 0.8 M Lithium sulfate monohydrate, 0.1 M Sodium acetate trihydrate pH 4.0, 4% v/v Polyethylene glycol 200 2. 0.2 M Lithium sulfate monohydrate, 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 26% v/v Polyethylene glycol 200 3. 0.05 M Calcium chloride dihydrate, 0.1 M MES monohydrate pH 6.0, 45% v/v Polyethylene glycol 200 4. 28% v/v 2-Propanol, 0.1 M BIS-TRIS pH 6.5, 3% v/v Polyethylene glycol 200

5. 20% v/v Tacsimate pH 7.0, 0.1 M HEPES pH 7.5, 2% v/v Polyethylene glycol 200

6. 10% v/v 2-Propanol, 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 26% v/v Polyethylene glycol 400 7. 0.2 M Ammonium acetate, 0.1 M Sodium citrate tribasic dihydrate pH 5.5, 24% v/v Polyethylene glycol 400 8. 0.2 M Ammonium sulfate, 0.1 M BIS-TRIS pH 6.5, 18% v/v Polyethylene glycol 400

9. 0.19 mM CYMAL®-7, 0.1 M HEPES pH 7.5, 40% v/v Polyethylene glycol 400

10. 6% v/v 2-Propanol, 0.1 M Sodium acetate trihydrate pH 4.5, 26% v/v Polyethylene glycol monomethyl ether 550 11. 1.8 M Ammonium sulfate, 0.1 M BIS-TRIS pH 6.5, 2% v/v Polyethylene glycol monomethyl ether 550 12. 0.15 M DL-Malic acid pH 7.0, 0.1 M Imidazole pH 7.0, 22% v/v Polyethylene glycol monomethyl ether 550 13. 0.1 M Succinic acid pH 7.0, 0.1 M BICINE pH 8.5, 30% v/v Polyethylene glycol monomethyl ether 550 14. 0.1 M Lithium sulfate monohydrate, 0.1 M Sodium citrate tribasic dihydrate pH 5.5, 20% w/v Polyethylene glycol 1,000 15. 0.1 M Sodium malonate pH 8.0, 0.1 M Tris pH 8.0, 30% w/v Polyethylene glycol 1,000

16. 4% v/v (+/-)-2-Methyl-2,4-pentanediol, 0.1 M Citric acid pH 3.5, 20% w/v Polyethylene glycol 1,500 17. 0.2 M L-Proline, 0.1 M HEPES pH 7.5, 24% w/v Polyethylene glycol 1,500

18. 10% v/v 2-Propanol, 0.1 M BICINE pH 8.5, 30% w/v Polyethylene glycol 1,500 19. 0.1 M Sodium chloride, 0.1 M BIS-TRIS propane pH 9.0, 25% w/v Polyethylene glycol 1,500

20. 0.02 M Nickel(II) chloride hexahydrate, 0.02 M Magnesium chloride hexahydrate, 0.02 M Cadmium chloride hydrate, 0.1 M Sodium acetate trihydrate pH 4.5, 24% w/v Polyethylene glycol monomethyl ether 2,000

21. 20% v/v 2-Propanol, 0.1 M MES monohydrate pH 6.0, 20% w/v Polyethylene glycol monomethyl ether 2,000 22. 0.2 M Ammonium citrate tribasic pH 7.0, 0.1 M Imidazole pH 7.0, 20% w/v Polyethylene glycol monomethyl ether 2,000 23. 4.0 M Potassium formate, 0.1 M BIS-TRIS propane pH 9.0, 2% w/v Polyethylene glycol monomethyl ether 2,000 24. 50% v/v Tacsimate pH 4.0, 0.1 M Sodium acetate trihydrate pH 4.5, 1% w/v Polyethylene glycol 3,350 25. 0.10% w/v n-Octyl-b-D-glucoside, 0.1 M Sodium citrate tribasic dihydrate pH 5.5, 22% w/v Polyethylene glycol 3,350 26. 2% v/v Tacsimate pH 7.0, 5% v/v 2-Propanol, 0.1 M Imidazole pH 7.0, 8% w/v Polyethylene glycol 3,350 27. 2% v/v 1,4-Dioxane, 0.1 M Tris pH 8.0, 15% w/v Polyethylene glycol 3,350

28. 18% v/v 2-Propanol, 0.1 M Sodium citrate tribasic dihydrate pH 5.5, 20% w/v Polyethylene glycol 4,000 29. 6% v/v Tacsimate pH 6.0, 0.1 M MES monohydrate pH 6.0, 25% w/v Polyethylene glycol 4,000 30. 0.2 M Magnesium formate dihydrate, 0.1 M Sodium acetate trihydrate pH 4.0, 18% w/v Polyethylene glycol monomethyl ether 5,000

31. 2% v/v Polyethylene glycol 400, 0.1 M Imidazole pH 7.0, 24% w/v Polyethylene glycol monomethyl ether 5,000 32. 0.2 M Sodium formate, 0.1 M BICINE pH 8.5, 20% w/v Polyethylene glycol monomethyl ether 5,000 33. 4% v/v 2-Propanol, 0.1 M BIS-TRIS propane pH 9.0, 20% w/v Polyethylene glycol monomethyl ether 5,000 34. 6% v/v Ethylene glycol, 0.1 M Citric acid pH 3.5, 10% w/v Polyethylene glycol 6,000

35. 0.15 M Lithium sulfate monohydrate, 0.1 M Citric acid pH 3.5, 18% w/v Polyethylene glycol 6,000 36. 10% v/v 2-Propanol, 0.1 M Sodium acetate trihydrate pH 4.0, 22% w/v Polyethylene glycol 6,000 37. 0.2 M Sodium chloride, 0.1 M Sodium acetate trihydrate pH 4.0, 22% w/v Polyethylene glycol 8,000 38. 20% v/v 2-Propanol, 0.1 M Tris pH 8.0, 5% w/v Polyethylene glycol 8,000

39. 10% v/v Polyethylene glycol 200, 0.1 M BIS-TRIS propane pH 9.0, 18% w/v Polyethylene glycol 8,000 40. 15% v/v 2-Propanol, 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 10% w/v Polyethylene glycol 10,000 41. 0.4 M Sodium malonate pH 6.0, 0.1 M MES monohydrate pH 6.0, 0.5% w/v Polyethylene glycol 10,000 42. 0.2 M Potassium sodium tartrate tetrahydrate, 0.1 M BIS-TRIS pH 6.5, 10% w/v Polyethylene glycol 10,000 43. 5% v/v (+/-)-2-Methyl-2,4-pentanediol, 0.1 M HEPES pH 7.5, 10% w/v Polyethylene glycol 10,000 44. 0.2 M Ammonium acetate, 0.1 M Tris pH 8.0, 16% w/v Polyethylene glycol 10,000 45. 5% v/v 2-Propanol, 0.1 M Citric acid pH 3.5, 6% w/v Polyethylene glycol 20,000

46. 1.0 M Sodium malonate pH 5.0, 0.1 M Sodium acetate trihydrate pH 4.5, 2% w/v Polyethylene glycol 20,000 47. 0.2 M Magnesium chloride hexahydrate, 0.1 M Sodium citrate tribasic dihydrate pH 5.0, 10% w/v Polyethylene glycol 20,000 48. 3% w/v Dextran sulfate sodium salt, 0.1 M BICINE pH 8.5, 15% w/v Polyethylene glycol 20,000

P e G r x 2 ™ f o r m u l a t i o n p o l y m e r Range from 3.5 to 9 BICINE BIS-TRIS BIS-TRIS propane Citric acid HEPES Imidazole MES monohydrate Sodium acetate Sodium citrate Tris Jeffamine® ED-2001 Jeffamine® M-600 ® Polyethylene glycol 200 Polyethylene glycol 300 Polyethylene glycol 400 Polyethylene glycol 1,000 Polyethylene glycol 1,500 Polyethylene glycol 3,350 Polyethylene glycol 4,000 Polyethylene glycol 6,000 Polyethylene glycol 8,000 Polyethylene glycol 10,000 Polyethylene glycol 20,000 Polyethylene glycol MME 550 Polyethylene glycol MME 2,000 Polyethylene glycol MME 5,000

pH b u f f e r s s e c o n d a r y r e a g e n t s b u f f e r s p o l y m e r s BICINE BIS-TRIS BIS-TRIS propane Citric acid HEPES Imidazole MES monohydrate Sodium acetate Sodium citrate Tris Range from 3.5 to 9 pH Polyethylene glycol 200 Polyethylene glycol 400 Polyethylene glycol 1,000 Polyethylene glycol 1,500 Polyethylene glycol 3,350 Polyethylene glycol 4,000 Polyethylene glycol 6,000 Polyethylene glycol 8,000 Polyethylene glycol 10,000 Polyethylene glycol 20,000 Polyethylene glycol MME 550 Polyethylene glycol MME 2,000 Polyethylene glycol MME 5,000 Additive Cation Detergent Organic Acid Polyol Volatile Organic

p e g r x

1 f a c t o r s

p e g r x

2 f a c t o r s

(14)

n Primary or secondary, polymer, salt and pH

matrix crystallization screen for biological macromolecules

n PEG/Ion is a sparse matrix profile of anions

and cations in the presence of monodisperse Polyethylene glycol 3,350 over pH 4.5 - 9.2

n PEG/Ion 2 screens a complete profile of

titrated organic acids at varying pH levels (3.7 - 8.8) in the presence of monodisperse PEG 3,350

n PEG/Ion HT combines PEG/Ion and PEG/Ion

2 in a single 96 Deep Well block

f e a t u r e s

PEG/Ion, developed by Hampton Research, is a crystallization screen designed to evalu-ate monodisperse, high purity Polyethylene glycol 3,350 and 48 unique salts represent-ing a very complete range of anions and cations frequently used in the crystallization of biological macromolecules. The primary screening variables are PEG, ion type, ionic strength, and pH. More than 60% of the published crystallizations utilized PEG as a primary crystallization reagent and in

approx-imately 50% of those reports, the PEG was combined with an ion as a secondary crystallization reagent. PEG/Ion 2 is an extension to the fundamental crystallization strategy in PEG/Ion. PEG/Ion 2 reagents cover the monodisperse, high purity Polyethylene glycol 3,350 and an array of neutralized and pH adjusted organic acids, multivalent ions, a novel Citrate BIS-TRIS propane buffer system and pH (4 - 8.8). The formulation of PEG/Ion 2 was developed at Hampton Research. Each of the 48 reagents in PEG/Ion 2 contains PEG 3,350 as the polymer (precipitant). The concentration of PEG is varied from 12% w/v to 20% w/v depending upon the type and concentration of buffer/salt paired with the polymer. Thirteen of the forty-eight PEG/Ion 2 reagents contain a separate buffer component. The remaining PEG/Ion 2 reagents are buffered by the titrated organic acid salt. Six of these thirteen conditions feature a novel Citric acid BIS-TRIS propane (CBTP) buffer. The CBTP buffer uses Citric acid and BIS-BIS-TRIS propane as the acid base pair to create a two component buffer system effective across pH 2.5 to 9.5. The ratio of Citric acid to BIS-TRIS propane determines the solution pH. Thirty-five of the forty-eight PEG/Ion 2 reagents contain a neutralized or pH adjusted organic acid in the presence of the polymer. Neutralized organic acids are highly effective

crystallization salts.1_{Four PEG/Ion 2 reagents feature polyvalent cations. Two of these reagents contain}

cation mixes, saving sample by screening six different cations with only two reagents. Tryptone, a casein digest combinatorial library of peptides, is included in PEG/Ion 2.

PEG/Ion contains 48 unique reagents, 10 ml each. PEG/Ion 2 contains 48 unique reagents, 10 ml each.

PEG/Ion HT contains 1 ml of each reagent from PEG/Ion and PEG/Ion 2 in a single Deep Well block format.

PEG/Ion 2

Measured pH range of kit is 3.7 to 8.8 at 25°C Average measured pH of kit is 6.4 at 25°C Median measured pH of kit is 6.7 at 25°C Mode measured pH of kit is 6.7 at 25°C

References

1. A comparison of salts for the crystallization of macromolecules. Alexander McPherson. Protein Science (2001), 10:418-422.

2. Searching for silver bullets: An alternative strategy for crystallizing macromolecules. Alexander McPherson and Bob Cudney. Journal of Structural Biology Volume 156, Issue 3 , December 2006, Pages 387-406

PEG/Ion Screen and PEG/Ion 2 Screen kits each contain 48 unique reagents. To order individual reagents, use Custom Shop™_{catalog numbers listed below. Refer to page 36 for further details.}

HR2-126 PEG/Ion Screen 10 ml, tube format $285.00

HR2-098 PEG/Ion 2 Screen 10 ml, tube format $285.00

HR2-139 PEG/Ion HT 1 ml, Deep Well block format $185.00

HR2-922-** PEG/Ion Screen Custom Shop 185 ml $138.00

HR2-998-** PEG/Ion 2 Screen Custom Shop 185 ml $138.00

screens

Crystals of Proteinase K. Preliminary crystallization con-ditions obtained using PEG/Ion Screen from Hampton Research.

Crystals grown at Hampton Research.

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screens A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 A11 A12 B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 E1 E2 E3 E4 E5 E6 E7 E8 E9 E10 E11 E12 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12 H1 H2 H3 H4 H5 H6 H7 H8 H9 H10 H11 H12 P e G / i o n s c r e e n ™ f o r m u l a t i o n

1. 0.2 M Sodium fluoride, 20% w/v Polyethylene glycol 3,350 2. 0.2 M Potassium fluoride, 20% w/v Polyethylene glycol 3,350 3. 0.2 M Ammonium fluoride, 20% w/v Polyethylene glycol 3,350 4. 0.2 M Lithium chloride, 20% w/v Polyethylene glycol 3,350

5. 0.2 M Magnesium chloride hexahydrate, 20% w/v Polyethylene glycol 3,350 6. 0.2 M Sodium chloride, 20% w/v Polyethylene glycol 3,350

7. 0.2 M Calcium chloride dihydrate, 20% w/v Polyethylene glycol 3,350 8. 0.2 M Potassium chloride, 20% w/v Polyethylene glycol 3,350 9. 0.2 M Ammonium chloride, 20% w/v Polyethylene glycol 3,350 10. 0.2 M Sodium iodide, 20% w/v Polyethylene glycol 3,350 11. 0.2 M Potassium iodide, 20% w/v Polyethylene glycol 3,350 12. 0.2 M Ammonium iodide, 20% w/v Polyethylene glycol 3,350 13. 0.2 M Sodium thiocyanate, 20% w/v Polyethylene glycol 3,350 14. 0.2 M Potassium thiocyanate, 20% w/v Polyethylene glycol 3,350 15. 0.2 M Lithium nitrate, 20% w/v Polyethylene glycol 3,350

16. 0.2 M Magnesium nitrate hexahydrate, 20% w/v Polyethylene glycol 3,350 17. 0.2 M Sodium nitrate, 20% w/v Polyethylene glycol 3,350

18. 0.2 M Potassium nitrate, 20% w/v Polyethylene glycol 3,350 19. 0.2 M Ammonium nitrate, 20% w/v Polyethylene glycol 3,350 20. 0.2 M Magnesium formate dihydrate, 20% w/v Polyethylene glycol 3,350 21. 0.2 M Sodium formate, 20% w/v Polyethylene glycol 3,350 22. 0.2 M Potassium formate, 20% w/v Polyethylene glycol 3,350 23. 0.2 M Ammonium formate, 20% w/v Polyethylene glycol 3,350 24. 0.2 M Lithium acetate dihydrate, 20% w/v Polyethylene glycol 3,350 25. 0.2 M Magnesium acetate tetrahydrate, 20% w/v Polyethylene glycol 3,350 26. 0.2 M Zinc acetate dihydrate, 20% w/v Polyethylene glycol 3,350 27. 0.2 M Sodium acetate trihydrate, 20% w/v Polyethylene glycol 3,350 28. 0.2 M Calcium acetate hydrate, 20% w/v Polyethylene glycol 3,350 29. 0.2 M Potassium acetate, 20% w/v Polyethylene glycol 3,350 30. 0.2 M Ammonium acetate, 20% w/v Polyethylene glycol 3,350 31. 0.2 M Lithium sulfate monohydrate, 20% w/v Polyethylene glycol 3,350 32. 0.2 M Magnesium sulfate heptahydrate, 20% w/v Polyethylene glycol 3,350 33. 0.2 M Sodium sulfate decahydrate, 20% w/v Polyethylene glycol 3,350 34. 0.2 M Potassium sulfate, 20% w/v Polyethylene glycol 3,350 35. 0.2 M Ammonium sulfate, 20% w/v Polyethylene glycol 3,350 36. 0.2 M Sodium tartrate dibasic dihydrate, 20% w/v Polyethylene glycol 3,350 37. 0.2 M Potassium sodium tartrate tetrahydrate, 20% w/v Polyethylene glycol 3,350 38. 0.2 M Ammonium tartrate dibasic, 20% w/v Polyethylene glycol 3,350

39. 0.2 M Sodium phosphate monobasic monohydrate, 20% w/v Polyethylene glycol 3,350 40. 0.2 M Sodium phosphate dibasic dihydrate, 20% w/v Polyethylene glycol 3,350 41. 0.2 M Potassium phosphate monobasic, 20% w/v Polyethylene glycol 3,350 42. 0.2 M Potassium phosphate dibasic, 20% w/v Polyethylene glycol 3,350 43. 0.2 M Ammonium phosphate monobasic, 20% w/v Polyethylene glycol 3,350 44. 0.2 M Ammonium phosphate dibasic, 20% w/v Polyethylene glycol 3,350 45. 0.2 M Lithium citrate tribasic tetrahydrate, 20% w/v Polyethylene glycol 3,350 46. 0.2 M Sodium citrate tribasic dihydrate, 20% w/v Polyethylene glycol 3,350 47. 0.2 M Potassium citrate tribasic monohydrate, 20% w/v Polyethylene glycol 3,350 48. 0.2 M Ammonium citrate dibasic, 20% w/v Polyethylene glycol 3,350

1. 0.1 M Sodium malonate pH 4.0, 12% w/v Polyethylene glycol 3,350 2. 0.2 M Sodium malonate pH 4.0, 20% w/v Polyethylene glycol 3,350 3. 0.1 M Sodium malonate pH 5.0, 12% w/v Polyethylene glycol 3,350 4. 0.2 M Sodium malonate pH 5.0, 20% w/v Polyethylene glycol 3,350 5. 0.1 M Sodium malonate pH 6.0, 12% w/v Polyethylene glycol 3,350 6. 0.2 M Sodium malonate pH 6.0, 20% w/v Polyethylene glycol 3,350 7. 0.1 M Sodium malonate pH 7.0, 12% w/v Polyethylene glycol 3,350 8. 0.2 M Sodium malonate pH 7.0, 20% w/v Polyethylene glycol 3,350 9. 4% v/v Tacsimate pH 4.0, 12% w/v Polyethylene glycol 3,350 10. 8% v/v Tacsimate pH 4.0, 20% w/v Polyethylene glycol 3,350 11. 4% v/v Tacsimate pH 5.0, 12% w/v Polyethylene glycol 3,350 12. 8% v/v Tacsimate pH 5.0, 20% w/v Polyethylene glycol 3,350 13. 4% v/v Tacsimate pH 6.0, 12% w/v Polyethylene glycol 3,350 14. 8% v/v Tacsimate pH 6.0, 20% w/v Polyethylene glycol 3,350 15. 4% v/v Tacsimate pH 7.0, 12% w/v Polyethylene glycol 3,350 16. 8% v/v Tacsimate pH 7.0, 20% w/v Polyethylene glycol 3,350 17. 4% v/v Tacsimate pH 8.0, 12% w/v Polyethylene glycol 3,350 18. 8% v/v Tacsimate pH 8.0, 20% w/v Polyethylene glycol 3,350 19. 0.1 M Succinic acid pH 7.0, 12% w/v Polyethylene glycol 3,350 20. 0.2 M Succinic acid pH 7.0, 20% w/v Polyethylene glycol 3,350 21. 0.1 M Ammonium citrate tribasic pH 7.0, 12% w/v Polyethylene glycol 3,350 22. 0.2 M Ammonium citrate tribasic pH 7.0, 20% w/v Polyethylene glycol 3,350 23. 0.1 M DL-Malic acid pH 7.0, 12% w/v Polyethylene glycol 3,350 24. 0.2 M DL-Malic acid pH 7.0, 20% w/v Polyethylene glycol 3,350 25. 0.1 M Sodium acetate trihydrate pH 7.0, 12% w/v Polyethylene glycol 3,350 26. 0.2 M Sodium acetate trihydrate pH 7.0, 20% w/v Polyethylene glycol 3,350 27. 0.1 M Sodium formate pH 7.0, 12% w/v Polyethylene glycol 3,350 28. 0.2 M Sodium formate pH 7.0, 20% w/v Polyethylene glycol 3,350 29. 0.1 M Ammonium tartrate dibasic pH 7.0, 12% w/v Polyethylene glycol 3,350 30. 0.2 M Ammonium tartrate dibasic pH 7.0, 20% w/v Polyethylene glycol 3,350

31. 2% v/v Tacsimate pH 4.0, 0.1 M Sodium acetate trihydrate pH 4.6, 16% w/v Polyethylene glycol 3,350 32. 2% v/v Tacsimate pH 5.0, 0.1 M Sodium citrate tribasic dihydrate pH 5.6, 16% w/v Polyethylene glycol 3,350 33. 2% v/v Tacsimate pH 6.0, 0.1 M BIS-TRIS pH 6.5, 20% w/v Polyethylene glycol 3,350

34. 2% v/v Tacsimate pH 7.0, 0.1 M HEPES pH 7.5, 20% w/v Polyethylene glycol 3,350 35. 2% v/v Tacsimate pH 8.0, 0.1 M Tris pH 8.5, 16% w/v Polyethylene glycol 3,350 36. (0.07 M Citric acid, 0.03 M BIS-TRIS propane / pH 3.4), 16% w/v Polyethylene glycol 3,350 37. (0.06 M Citric acid, 0.04 M BIS-TRIS propane / pH 4.1), 16% w/v Polyethylene glycol 3,350 38. (0.05 M Citric acid, 0.05 M BIS-TRIS propane / pH 5.0), 16% w/v Polyethylene glycol 3,350 39. (0.04 M Citric acid, 0.06 M BIS-TRIS propane / pH 6.4), 20% w/v Polyethylene glycol 3,350 40. (0.03 M Citric acid, 0.07 M BIS-TRIS propane / pH 7.6), 20% w/v Polyethylene glycol 3,350 41. (0.02 M Citric acid, 0.08 M BIS-TRIS propane / pH 8.8), 16% w/v Polyethylene glycol 3,350 42. 0.02 M Calcium chloride dihydrate, 0.02 M Cadmium chloride hydrate,

0.02 M Cobalt(II) chloride hexahydrate, 20% w/v Polyethylene glycol 3,350 43. 0.01 M Magnesium chloride hexahydrate, 0.005 M Nickel(II) chloride hexahydrate 0.1 M HEPES sodium pH 7.0, 15% w/v Polyethylene glycol 3,350 44. 0.02 M Zinc chloride, 20% w/v Polyethylene glycol 3,350 45. 0.15 M Cesium chloride, 15% w/v Polyethylene glycol 3,350 46. 0.2 M Sodium bromide, 20% w/v Polyethylene glycol 3,350

47. 1% w/v Tryptone, 0.05 M HEPES sodium pH 7.0, 12% w/v Polyethylene glycol 3,350 48. 1% w/v Tryptone, 0.05 M HEPES sodium pH 7.0, 20% w/v Polyethylene glycol 3,350

P e G / i o n 2 s c r e e n ™ f o r m u l a t i o n

m u l t i v a l e n t i o n s

b u f f e r s

o r g a n i c s a l t s

Neutralized Organic Salts BIS-TRIS Citric acid BIS-TRIS propane (CBTP)

HEPES HEPES sodium Sodium acetate Sodium citrate Tris Range from 4 to 8 pH Ammonium tartrate Ammonium citrate Malic acid Succinic acid Sodium acetate Sodium formate Sodium malonate Tacsimate™ Range from 4.0 to 9.0 Polyethylene glycol 3,350 Acetate Citrate Chloride Fluoride Formate Iodide Nitrate Phosphate Sulfate Thiocyanate Tartrate pH a n i o n p o l y m e r Ammonium_Calcium Lithium Magnesium Potassium Sodium Zinc c a t i o n Polyethylene glycol 3,350 p o l y m e r Cadmium chloride Calcium chloride Cesium chloride Cobalt(II) chloride Magnesium chloride Nickel(II) chloride Sodium bromide Zinc chloride p e p t i d e s / a m i n o a c i d s Tryptone

p e g / i o n

f a c t o r s

p e g / i o n

2 f a c t o r s

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screens

Grid Screen Salt HT™ combines Grid Screen Ammonium Sulfate, Grid Screen Sodium Malonate, Quik Screen and Grid Screen Sodium Chloride into a single 96 Deep Well block format. The block contains 1 ml of each reagent.

Grid Screen Ammonium Sulfate systemati-cally evaluates Ammonium sulfate at four concentrations (0.8, 1.6, 2.4, 3.2 M) versus 6 pH levels (4, 5, 6, 7, 8, 9).

Grid Screen Sodium Malonate evaluates Sodium malonate at six concentrations (1.0, 1.5, 1.9, 2.4, 2.9, 3.4 M) versus four pH levels (4, 5, 6, 7).

Quik Screen evaluates Sodium potassium phosphate at four concentrations (0.8, 1.0, 1.4, 1.8 M) versus six pH levels (5.0, 5.6, 6.3, 6.9, 7.5, 8.2).

Grid Screen Sodium Chloride evaluates Sodium chloride at four concentrations (1.0, 2.0, 3.0, 4.0 M) versus six pH levels (4, 5, 6, 7, 8, 9).

Grid Screen PEG 6000 evaluates Polyethylene glycol at four concentrations (5, 10, 20, 30 %w/v) versus six pH levels (4, 5, 6, 7, 8, 9).

Grid Screen MPD evaluates MPD at four concentrations (10, 20, 40, 65 %v/v) versus six pH levels (4, 5, 6, 7, 8, 9).

Grid Screen PEG/LiCl evaluates Polyethylene glycol at four concentrations (0, 10, 20, 30 %w/v) in the presence of 1.0 M Lithium chloride versus six pH levels (4, 5, 6, 7, 8, 9).

Grid Screens and Quik Screen each contain 10 ml of 24 unique reagents.

Ready-to-use reagents are sterile filtered and formulated with ultra-pure Type 1 water, using the high-est purity salts, polymers, organics and buffers.

References

1. A protein crystallization strategy using automated grid searches on successively finer grids. Patricia C. Weber. Methods: A Companion to Methods in Enzymology Vol. 1, No. 1, August, pp. 31-37, 1990.

2. Protein Crystallization; Techniques, Strategies, and Tips. A Laboratory Manual. Edited by Terese M. Bergfors. International University Line, 1999. ISBN 0-9636817-5-3.

f e a t u r e s

n Primary or secondary, salt, polymer, organic

and pH grid crystallization screen for biologi-cal macromolecules

n A systematic grid screen varying salt or

poly-mer or organic versus pH

n Samples pH 4 to 9

n Combine reagents within or between screens

to customize your screen

n Combine reagents between rows or columns

to create expanded grid screens

Each Grid Screen Salt HT kit contains 96 unique reagents.

HR2-248 Grid Screen Salt HT 1 ml, Deep Well block format $185.00 Number of crystals grown

versus varying A/S concentrations [M] Ammonium Sulfate [M] 60 50 40 30 20 10 0 0 10 20 30 40 50 60 0 to 5 6 to 10 11 to 16 16 to 20 21 to 25 25 to 30 30+ Crystals Grown Number of Cr ystal s % w/v PEG 6000

Number of crystals grown versus varying PEG 6000

concentrations (% w/v) 70 to 79 60 to 69 50 to 59 40 to 49 30 to 39 20 to 29 10 to 19 0 to 9 Crystals Grow n 30 25 20 15 10 5 0 Number of Crystals % v/v MPD 0 5 10 15 20 25 30

Number of crystals grown versus varying MPD concentrations (% v/v)

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screens

HR2-211 Grid Screen Ammonium Sulfate 10 ml, tube format $175.00

HR2-924-** Grid Screen Ammonium Sulfate Custom Shop 185 ml $138.00

** = reagent number A1-D6 Order Information

Each Grid Screen and Quik Screen kit contains 24 unique reagents. To order individual reagents, use Custom Shop™ catalog number listed below. Refer to page 36 for further details.

HR2-247 Grid Screen Sodium Malonate 10 ml, tube format $175.00

HR2-947-** Grid Screen Sodium Malonate Custom Shop 185 ml $138.00

** = reagent number A1-D6

HR2-221 Quik Screen 10 ml, tube format $175.00

HR2-921-** Quik Screen Custom Shop 185 ml $138.00

HR2-219 Grid Screen Sodium Chloride 10 ml, tube format $175.00

HR2-932-** Grid Screen Sodium Chloride Custom Shop 185 ml $138.00

HR2-213 Grid Screen PEG 6000 10 ml, tube format $175.00

HR2-926-** Grid Screen PEG 6000 Custom Shop 185 ml $138.00

HR2-215 Grid Screen MPD 10 ml, tube format $175.00

HR2-930-** Grid Screen MPD Custom Shop 185 ml $138.00

HR2-217 Grid Screen PEG/LiCl 10 ml, tube format $175.00

HR2-928-** Grid Screen PEG/LiCl Custom Shop 185 ml $138.00

[ M ] Sodium malonate 1 1.0 2 1.5 3 1.9 4 2.4 5 2.9 6 3.4 B A C 5.0 4.0 6.0 7.0 D pH [ M ] Sodium/Potassium Phosphate 1 5.0 2 5.6 3 6.3 4 6.9 5 7.5 6 8.2 B A C 1.0 M 0.8 M 1.4 M 1.8 M D pH 0.1 M Buffer [ M ] Ammonium sulfate 1 4 2 5 3 6 4 7 5 8 6 9 1.6 M B 0.8 M A 2.4 M C 3.0 M D Citri c aci d MES HEPES Tr is BICINE Citri c aci d [ % w/v ] Polyethylene glycol 6,000 0.1 M Buffer 1 4 2 5 3 6 4 7 5 8 6 9 10% B 5% A 20% C 30% D Citri c aci d MES HEPES Tri s BICINE Citri c aci d [ M ] Sodium chloride 0.1 M Buffer 1 4 2 5 3 6 4 7 5 8 6 9 2.0 M B 1.0 M A 3.0 M C 4.0 M D Citri c aci d MES HEPES Tri s BICINE Citri c aci d [ % v/v ] MPD 0.1 M Buffer 1 4 2 5 3 6 4 7 5 8 6 9 20% B 10% A 40% C 65% D Citri c aci d MES HEPES Tri s BICINE Citri c aci d [ % w/v ] Polyethylene glycol 6,000 1.0 M Lithium chloride in all reagents

0.1 M Buffer 1 4 2 5 3 6 4 7 5 8 6 9 10% B 5% A 20% C 30% D Citri c aci d MES HEPES Tr is BICINE Citri c aci d

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Crystals grown using SaltRx, reagent 62. They were grown using the Cyberlab C240 robot and the Neuro Probe plate. These were grown just in time to freeze and take to the synchrotron (no time for optimizations). MAD data were collected on these crystals and the structure has been solved to 2.3Å resolution.

Courtesy of Annie Hassell.

GlaxoSmithKline

s u c c e s s s t o r y

n Primary or secondary, salt and pH matrix

crystallization screen for biological macromol-ecules

n Salt versus pH matrix crystallization screen

n Samples pH 4 - 9

n 22 unique salts versus salt concentration

and pH

n Compatible with microbatch, vapor diffusion,

liquid & gel diffusion methods

n Preformulated, ready to screen

n All salts and buffers in screen readily

avail-able as Optimize reagents for reproducing and optimizing crystals

f e a t u r e s

SaltRx was developed by Hampton Research as a primary and secondary, salt and pH based crystallization screen for biological macromolecules. Salt is the only primary crystallization reagent (precipitant) utilized. Based on a design of 96 conditions, the screen evaluates a broad portfolio of crystalli-zation salts of varying concentration and pH. The selection, concentration, and pH of the

salts were determined by data mining the BMCD1_{and other crystallization databases, crystallization reports}

in the literature, and internal crystallization trials performed at Hampton Research. Based on this analysis, up to 35% of protein crystallizations involve salt as the primary crystallization reagent.

SaltRx can be used as a primary crystallization screen when salt, ionic strength and pH are desired or suspected as appropriate crystallization variables. It is also useful as a secondary screen when salt based reagents/conditions from screens such as Index, Crystal Screen, and Grid Screen produce crystals and when further screening for additional salt conditions or optimization is desired.

SaltRx is designed to be used as a 96 reagent screen. SaltRx 1 contains 48 unique reagents, 10 ml each. SaltRx 2 contains 48 unique reagents, 10 ml each.

SaltRx HT contains 1 ml of each reagent from SaltRx 1 and SaltRx 2 in a single Deep Well block format. Ready-to-use reagents are sterile filtered and formulated with ultra-pure Type 1 water, using the highest purity salts, polymers, organics and buffers.

References

1. Gilliland, G.L., Tung, M., Blakeslee, D.M. and Ladner, J. 1994. The Biological Macromolecule Crystallization Database, Version 3.0: New Features, Data, and the NASA Archive for Protein Crystal Growth Data. Acta Crystallogr. D50 408-413.

2. Crystallization and preliminary crystallographic analysis of molybdenumcofactor biosynthesis protein C from Thermus thermophilus. S. P. Kanaujia, C. V. Ranjani, J. Jeyakanthan, S. Baba, L. Chen, Z.-J. Liu, B.-C. Wang, M. Nishida, A. Ebihara, A. Shinkai, S. Kuramitsu, Y. Shiro,

K. Sekar and S. Yokoyama. Acta Cryst. (2007). F63, 27-29.

SaltRx 1 and SaltRx 2 kits each contain 48 unique reagents. To order individual reagents, use Custom Shop™_{catalog number listed below. Refer to page 36 for further details.}

HR2-107 SaltRx 1 10 ml, tube format $285.00

HR2-109 SaltRx 2 10 ml, tube format $285.00

HR2-136 SaltRx HT 1 ml, Deep Well block format $185.00

HR2-907-** SaltRx 1 Custom Shop 185 ml $138.00

HR2-909-** SaltRx 2 Custom Shop 185 ml $138.00